The chronic administration of many chemical agents results in the appearance of malignant tumors in target tissues. These malignancies are characterized by alterations in control of cellular functions which suggest that mechanisms of genetic modulation have been altered. Recent evidence has accumulated which suggests that non-histone chromosomal proteins (NHCP) are involved in transcriptional control. The object of this proposal is to analyze these NHCP in normal liver, at well delineated stages of carcinogenic evolution and in the tumors which arise. We will use our previously described, intermittent dietary regimen of N-2-Fluorenylacetamide (FAA) and examine liver exposed to subcarcinogenic doses, the premalignant nodules which arise with further exposure and subsequent tumors. Chromatin will be isolated, purified, fractionated into eu-and heterochromatin by controlled shearing, and separated in glycerol gradients. Transcriptional capacity will be monitored with E. coli polymerase. The NHCP of these fractions will be analyzed by Isoelectric focussing followed by two dimensional acrylamide electrophoresis. In addition to this quantitative and qualitative analysis of NHCP patterns, the binding of labelled FAA (and its metabolites) to selective protein species will be measured. Lastly, the nature of specific proteins responsible for binding of physiologic stimuli such as triiodothyronine, will be identified and examined in the above tissues as a "probe" for altered chromatin composition.